Youngeun Kim

Nattokinase improves blood flow by inhibiting platelet aggregation and thrombus formation

The effects of nattokinase on the in vitro platelet aggregation and in vivo thrombosis were investigated in comparison with aspirin. Rabbit platelet-rich plasma was incubated with nattokinase and aggregation inducers collagen and thrombin, and the platelet aggregation rate was analyzed. Nattokinase significantly inhibited both the collagen- and thrombin-induced platelet aggregations. Nattokinase also reduced thromboxane B2 formation from collagen-activated platelets in a concentration-dependent manner. Rats were orally administered with nattokinase for 1 week, and their carotid arteries were exposed. Arterial thrombosis was induced by applying 35% FeCl3-soaked filter paper for 10 min, and the blood flow was monitored with a laser Doppler probe. Nattokinase delayed the FeCl3-induced arterial occlusion in a dose-dependent manner, doubling the occlusion time at 160 mg/kg. In addition, a high dose (500 mg/kg) of nattokinase fully prevented the occlusion, as achieved with aspirin (30 mg/kg). The results indicate that nattokinase extracted from fermented soybean inhibit platelet aggregation by blocking thromboxane formation, and thereby delay thrombosis following oxidative arterial wall injury. Therefore, it is suggested that nattokinase could be a good candidate without adverse effects for the improvement of blood flow.

Inhibitory effect of punicalagin on lipopolysaccharide-induced neuroinflammation, oxidative stress and memory impairment via inhibition of nuclear factor-kappaB.

&NA; Neuroinflammation is significant in the pathogenesis and development of Alzheimers disease (AD). Previously, we showed lipopolysaccharide (LPS)‐induced neuroinflammation caused memory impairment. We investigated the possible preventive effects of punicalagin (PUN), a component of pomegranate, on memory deficiency caused by LPS, along with the fundamental mechanisms. LPS‐treated cultured astrocytes and microglial BV‐2 cells were investigated for anti‐neuroinflammatory effects of PUN. PUN (1.5 mg/kg) ameliorates LPS (250 &mgr;g/kg daily 7 times)‐induced memory impairment as well as prevents the LPS‐induced expression of inflammatory proteins. In in vitro study, we also found that PUN (1 &mgr;g/ml) inhibited the LPS‐(10, 20 and 50 &mgr;M) induced expression of iNOS and Cox‐2 as well as the production of ROS, NO, TNF‐&agr; and IL‐1&bgr;. PUN also suppress activation of NF‐&kgr;B via inhibition of I&kgr;B degradation as well as p50 and p65 translocation into the nucleus in LPS treated mouse brain and cultured astrocytes and microglial BV‐2 cells. Consistent with the inhibitory effect on neuro inflammation, PUN inhibited LPS‐induced A&bgr;1‐42 generation through down‐regulation of APP and BACE1 expression in in vivo and in vitro study. Moreover, PUN directly binds to NF‐&kgr;B subunit p50 evidenced by a docking model and pull down assay. These results suggest that PUN inhibits LPS‐induced memory impairment via anti‐inflammatory and anti‐amylogenic mechanisms through inhibition of NF‐&kgr;B activation. HighlightsNeuroinflammation and amyloidogenesis are main symptoms of Alzheimers disease.NF‐&kgr;B activation can induce the inflammation and amyloidogenesis pathways.Punicalagin inhibits NF‐&kgr;B activation through direct binding to its subunit P50.Punicalagin reduces LPS‐induced neuroinflammation and amyloidogenesis.Punicalagin is a possible candidate for treating Alzheimers disease.

Perilla oil improves blood flow through inhibition of platelet aggregation and thrombus formation.

The inhibitory effects of perilla oil on the platelet aggregation in vitro and thrombosis in vivo were investigated in comparison with aspirin, a well-known blood flow enhancer. Rabbit platelet-rich plasma was incubated with perilla oil and aggregation inducers collagen or thrombin, and the platelet aggregation rate was analyzed. Perilla oil significantly inhibited both the collagen- and thrombin-induced platelet aggregations, in which the thromboxane B2 formation from collagen-activated platelets were reduced in a concentration-dependent manner. Rats were administered once daily by gavage with perilla oil for 1 week, carotid arterial thrombosis was induced by applying 35% FeCl3-soaked filter paper for 10 min, and the blood flow was monitored with a laser Doppler probe. Perilla oil delayed the FeCl3-induced arterial occlusion in a dose-dependent manner, doubling the occlusion time at 0.5 mL/kg. In addition, a high dose (2 mL/kg) of perilla oil greatly prevented the occlusion, comparable to the effect of aspirin (30 mg/kg). The results indicate that perilla oil inhibit platelet aggregation by blocking thromboxane formation, and thereby delay thrombosis following oxidative arterial wall injury. Therefore, it is proposed that perilla oil could be a good candidate without adverse effects for the improvement of blood flow.

Parkin deficiency exacerbate ethanol-induced dopaminergic neurodegeneration by P38 pathway dependent inhibition of autophagy and mitochondrial function

Parkinsons disease (PD) is a neurodegenerative disease characterized by selective degeneration of dopaminergic neurons in the substantia nigra. Parkin (which encoded by Park2), an E3 ubiquitin ligase, is the most frequently mutated gene that has casually been linked to autosomal recessive early onset familial PD. We tested the effect of Park2 on ethanol-induced dopaminergic neurodegeneration in Park2 knockout (KO) transgenic mice after chronic ethanol feeding. Male Park2 wild type (WT) and KO mice (8 weeks old) were fed on a Lieber-DeCarli diet containing 6.6% ethanol for 2 weeks, and compared their responses. We found that knockout of Park2 exacerbates ethanol-induced behavioral impairment as well as dopamine depletion. In the mechanism study, we found that knockout of Park2 increased reactive oxygen species (ROS) production, mitophagy formation, mitochondrial dysfunction, and expression of pro-apoptotic proteins, but decreased expression of pro-autophagic proteins. Knockout of Park2 also increased ethanol-induced activation of p38 mitogen-activated protein kinase. In addition, ROS production, mitophagy formation, mitochondrial dysfunction, and expression of pro-apoptotic proteins were increased, but expression of pro-autophagic proteins were decreased by a treatment of ethanol (100 μM) in Park2 siRNA-transfacted PC12 cells (5 μM). Moreover, the exacerbating effects of Park2 deletion on ethanol-induced ROS generation, mitophagy, mitochondrial dysfunction as well as cell death were reduced by p38 specific inhibitor (SB203580) in in vitro (10 μM) and in vivo 10 mg/kg). Park2 deficiency exacerbates ethanol-induced dopaminergic neuron damage through p38 kinase dependent inhibition of autophagy and mitochondrial function.

Highly endoreduplicated floral organs of somaclonal variants in clonally propagated Phalaenopsis ‘Spring Dancer’

A micropropagation system is essential for obtaining elite clones of Phalaenopsis in the floricultural industry; however, somaclonal variation frequently occurs during long-term micropropagation, which reduces the uniformity and quality of clonally propagated plants. To detect the factors influencing this variation and thus obtain high-quality, uniform Phalaenopsis plants, we analyzed somaclonal variants in the Phalaenopsis ‘Spring Dancer’ (SD) variety collected from an orchid farm, which were grouped into three types according to flower morphology (SD-N, SD-V1, SD-V2). Morphological differences were observed between all SD groups; in particular, the diameter of the petal and thickness of each organ were significantly larger in the variants than SD-N (normal) plants. Both SD variant groups showed the same ploidy level as that of SD-N. The degree of endoreduplication of the petals and flower stalks was higher in the variants (4C–16C) than in SD-N (2C–8C). Cell area was increased and the shape of epidermal cells was changed in both variants. Expression levels of MADS box transcription factor (MADS4) of the flowers were higher than those of the leaves in the variants. Among the three flower organs, MADS4 expression was the highest in the petals of SD-V2. These results revealed that MADS4 is involved in the modification of flower morphology in somaclonal SD variants. High ploidy 2 (HPY2) expression was highest in the flowers of SD-V1, whereas its expression in the leaf was similar to that of SD-N. Among the various flower organs, HPY2 expression was highest in the lips of SD-V1.

Ethanol Extracts of Unroasted Coffea canephora Robusta Beans Suppress Adipogenesis in Preadipocytes and Fat Accumulation in Rats Fed a High-fat Diet

Anti-adipogenic and anti-obesity effects of a green coffee been extract (GCE) from unroasted Coffea canephora Robusta beans were investigated in vitro and in vivo. Insulin-induced adipogenesis in 3T3-L1 preadipocytes was inhibited by 25.6 and 87.9% following treatment with 100 and 500 μg/mL GCE, respectively. Expression of adipogenesis-specific genes was down-regulated. Body weight gains of C57BL/6 mice fed a high-fat diet (HFD) were suppressed in a concentration-dependent manner using GCE (0.3 or 1% in the diet), exhibiting a marked lower gain in 1% GCE-fed mice than in normal diet-fed animals. Decreases in body fat weights and the size of adipocytes were confirmed. GCE supplementation reduced concentrations of blood lipids and leptin, and attenuated hepatic steatosis. GCE inhibits adipogenesis by modulating cell signaling and improves dietary obesity by reducing lipid accumulation and the size of adipocytes.

Overexpression of SUMO E3 Ligase HPY2 Regulates the Cell Cycle in Petunia Development

Sumoylation is a post-translational modification that mediates several regulatory processes in plants. It is associated with developmental processes such as cellular division, expansion and differentiation, and vegetative and reproductive growth. The primary function of SUMO E3 ligases is to control the mitotic cycle and maintain meristematic cells. Arabidopsis thaliana HIGH PLOIDY2 (HPY2), a SUMO E3 ligase, might therefore be expected to promote cell division and increase organ size when overexpressed in the model ornamental plant Petunia multiflora. One month after Agrobacterium-mediated transformation, nine putative transgenic P. multiflora plants were regenerated on selection medium, and were found to have high expression levels of AtHPY2. The proportion of cells in G2M phase, length of in vitro-cultivated plantlets, and trichome length all increased in transgenic lines compared with the wild type (WT). Transgenic petunia lines also exhibited increased growth, leaf and flower area, and flower number. Flowering time was also faster in transgenic P. multiflora than in the WT, and flower numbers increased. These results suggest that AtHPY2 overexpression stimulates cell division. This finding may be useful in controlling plant and flower size, and flowering timing and abundance in P. multiflora and other ornamentals.